CN110483462A - Polyphenol compound and preparation method thereof with neuroprotective activity - Google Patents
Polyphenol compound and preparation method thereof with neuroprotective activity Download PDFInfo
- Publication number
- CN110483462A CN110483462A CN201910778418.8A CN201910778418A CN110483462A CN 110483462 A CN110483462 A CN 110483462A CN 201910778418 A CN201910778418 A CN 201910778418A CN 110483462 A CN110483462 A CN 110483462A
- Authority
- CN
- China
- Prior art keywords
- compound
- fraction
- preparation
- water
- gradient
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C67/00—Preparation of carboxylic acid esters
- C07C67/48—Separation; Purification; Stabilisation; Use of additives
- C07C67/56—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
- C07C69/76—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring
- C07C69/84—Esters of carboxylic acids having a carboxyl group bound to a carbon atom of a six-membered aromatic ring of monocyclic hydroxy carboxylic acids, the hydroxy groups and the carboxyl groups of which are bound to carbon atoms of a six-membered aromatic ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/06—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
- C07D311/20—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 hydrogenated in the hetero ring
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
- C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
- C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
- C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Pharmacology & Pharmacy (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention belongs to pharmaceutical technology field, it is related to polyphenol compound and preparation method thereof in pithecellobium clypearia, the invention further relates to application of the polyphenol compound in preparation neurodegenerative disease drug.The structure of the polyphenol compound is as follows, and is prepared via a method which: pithecellobium clypearia branches and leaves are extracted with ethyl alcohol, and extracting solution is concentrated under reduced pressure, and combined extract is condensed into medicinal extract.It is extracted with ethyl acetate, acetic acid ethyl ester extract is using isolated following 5 compounds of the means such as silica gel chromatographic column, HP20 chromatographic column, ODS chromatographic column, HPLC.The compound has anti-H2O2The neuroprotective activity of induction acts on.
Description
Technical field
The invention belongs to pharmaceutical technology field, it is related to polyphenol compound and preparation method thereof in pithecellobium clypearia, the present invention is also
It is related to application of the polyphenol compound in preparation neurodegenerative disease drug.
Background technique
Neurodegenerative disease (Neurodegenerative diseases) is with the slow and selective neuronal that breaks out
Dysfunction be characterized performance, the irreversible nerve of one kind due to caused by the neuronal cell loss of brain and spinal cord
Systemic disease mainly includes Alzheimer's disease (Alzheimer disease, AD), Parkinson's disease (Parkinson '
Sdisease, PD) and Huntington's disease (Huntington ' s disease, HD) etc..According to modern research shows that its pathogenesis
It may include following several respects: (1) inherent cause (2) protein abnormal stacking (3) oxidative stress (4) metal ion disorder (5)
DNA damage reparation (6) energy metabolism impairment etc..
Pithecellobium clypearia (Pithecellobium clypearia.) is that pulse family pithecellobium clypearia belongs to magaphanerophytes, is distributed in China, China
The ground such as south and Zhejiang, Fujian.Pithecellobium clypearia is first recorded in the Compendium of Material Medica of Li Shizhen of the Ming Dynasty, and drying stem and branch with leaf can be used as medicine, nature and flavor
Bitterness is cold, has effects that clearing heat and detoxicating, hygroscopic sore, cures mainly a variety of heat toxin syndromes.Mainly have in the patent medicine of clinical application at present
HOUERHUAN XIAOYAN JIAONANG, Ramulus Et Folium Pithecellobii Lucidi antiimflammatory tablet, pithecellobium clypearia antiphlogistic granules have the effect of clearing heat and detoxicating, cool blood detumescence, antidiarrheal,
Cure mainly the infection of the upper respiratory tract, acute laryngopharyngitis, acute tonsillitis, acute gastroenteritis, it can also be used to bacillary dysentery.Modern times grind
Study carefully discovery, flavonoids, Phenylpropanoid Glycosides class, organic phenolic acid class, triterpene and steroid compound are mainly contained in pithecellobium clypearia.Its modern medicine
It manages to study and is mostly focused on flavonoids, in terms of the bioactivity of organic liposoluble ingredient, there is anti-oxidant, anti-inflammatory, adjusting to be immunized,
The effects of antiviral, hypoglycemic.
Five polyphenol compounds involved in the present invention, the experiment proved that, the compound is protected with nerve cell
Shield activity can be applied to treatment neurodegenerative disease.
Summary of the invention
An object of the present invention is to provide five polyphenol compounds；
The second object of the present invention is to provide the preparation method of five polyphenol compounds；
The third object of the present invention is to provide five polyphenol compounds in preparation neurodegenerative disease drug
Using.
Five flavone compounds of the present invention, are respectively designated as: (1S)-pithecellobiumin C, (3S)-
pithecellobiumin D、pithecellobiumin E、pithecellobiumin F、pithecellobiumin G。
Chemical structural formula is as follows:
Five compounds of the present invention the preparation method is as follows:
Pithecellobium clypearia branches and leaves are extracted with ethyl alcohol, and extracting solution is concentrated under reduced pressure, and combined extract is condensed into medicinal extract.With acetic acid second
Ester is extracted, and acetic acid ethyl ester extract is separated using means such as silica gel chromatographic column, HP20 chromatographic column, ODS chromatographic column, HPLC
To above 5 compounds.
Specifically: five compounds process for production thereof are as follows:
(1) dry pithecellobium clypearia branches and leaves are taken, are extracted with 70-80% ethyl alcohol, extracting solution is obtained and is condensed into medicinal extract.Total medicinal extract
It after being suspended with water, is extracted with ethyl acetate, ethyl acetate layer uses silicagel column, carries out ladder with methylene chloride-methanol system
Degree elution, collects fraction A.
(2) fraction A is eluted through ODS column chromatography through ethanol-water system, collects fraction B.
(3) fraction B collects fraction C by methylene chloride-methanol system elutions through silicagel column.
(4) fraction C collects fraction D through HP20 resin with ethanol-water system gradient elution.
(5) fraction D collects fraction G, fraction H, fraction I through ethanol-water system elution, fraction J evaporates by ODS column respectively
Divide H mainly to collect in 40%-50% (40%, 45%, 50%) gradient, and is made in 30%-45% acetonitrile-water by HPLC method
It is standby to obtain compound 1, compound 2, compound 3, compound 4, compound 5.
Wherein:
The gradient of methylene chloride-methanol in step (1) are as follows: 50:1-1:2.
Fraction A is the eluate of methylene chloride-methanol 30:1-20:1 in step (1).
The gradient of alcohol-water is 30%-90% in step (2).
Fraction B is the eluate of alcohol-water 60% in step (2).
The gradient of methylene chloride-methanol in step (3) are as follows: 50:1-1:2.
Fraction C is the eluate of methylene chloride-methanol 20:1-10:1 in step (3).
The gradient of alcohol-water is 30%-90% in step (4).
Fraction D is the eluate of alcohol-water 50% in step (4).
The gradient of alcohol-water is 40%-70% in step (5).
In the present invention, five polyphenol compounds have also been carried out in vitro to by H2O2The neuroblastoma SH- of induction
The neuroprotection of SY5Y cellular damage measures, and compound 1, compound 2, compound 3, compound 4 are in difference as the result is shown
Concentration has the anti-H for being better than positive drug2O2The neuroprotective activity of induction, compound 5 have and the comparable anti-H of positive drug2O2It lures
The neuroprotective activity led.
Detailed description of the invention:
Fig. 1 is that the UV of compound 1 is composed；
Fig. 2 is that the IR of compound 1 is composed；
Fig. 3 is that the HRESIMS of compound 1 is composed；
Fig. 4 is compound 11H-NMR(400MHz,DMSO-d6) spectrum；
Fig. 5 is compound 113C-NMR(100MHz,DMSO-d6) spectrum；
Fig. 6 is HMQC (600MHz, the DMSO-d of compound 16) spectrum；
Fig. 7 is HMBC (600MHz, the DMSO-d of compound 16) spectrum；
Fig. 8 is that the UV of compound 2 is composed；
Fig. 9 is that the IR of compound 2 is composed；
Figure 10 is that the HRESIMS of compound 2 is composed；
Figure 11 is compound 21H-NMR(400MHz,CD4O it) composes；
Figure 12 is compound 213C-NMR(100MHz,CD4O it) composes；
Figure 13 is HMQC (600MHz, the CD of compound 24O it) composes；
Figure 14 is HMBC (600MHz, the CD of compound 24O it) composes；
Figure 15 is the ECD spectrum of compound 1 and compound 2；
Figure 16 is that the UV of compound 3 is composed；
Figure 17 is that the IR of compound 3 is composed；
Figure 18 is that the HRESIMS of compound 3 is composed；
Figure 19 is compound 31H-NMR(400MHz,DMSO-d6) spectrum；
Figure 20 is compound 313C-NMR(100MHz,DMSO-d6) spectrum；
Figure 21 is HMQC (600MHz, the DMSO-d of compound 36) spectrum；
Figure 22 is HMBC (600MHz, the DMSO-d of compound 36) spectrum；
Figure 23 is that the UV of compound 4 is composed；
Figure 24 is that the IR of compound 4 is composed；
Figure 25 is that the HRESIMS of compound 4 is composed；
Figure 26 is compound 41H-NMR(400MHz,DMSO-d6) spectrum；
Figure 27 is compound 413C-NMR(100MHz,DMSO-d6) spectrum；
Figure 28 is HMQC (600MHz, the DMSO-d of compound 46) spectrum；
Figure 29 is HMBC (600MHz, the DMSO-d of compound 46) spectrum；
Figure 30 is that the UV of compound 5 is composed；
Figure 31 is that the IR of compound 5 is composed；
Figure 32 is that the HRESIMS of compound 5 is composed；
Figure 33 is compound 51H-NMR(400MHz,DMSO-d6) spectrum；
Figure 34 is compound 513C-NMR(100MHz,DMSO-d6) spectrum；
Figure 35 is HMQC (600MHz, the DMSO-d of compound 56) spectrum；
Figure 36 is HMBC (600MHz, the DMSO-d of compound 56) spectrum.
Specific embodiment
The preparation method of five polyphenol compounds in 1 pithecellobium clypearia of embodiment
Dry pithecellobium clypearia branches and leaves 20kg is taken, 70% alcohol reflux measured with 6 times extracts 2 times, and 2 hours every time, merging mentioned
Liquid is taken, medicinal extract 0.9kg is concentrated under reduced pressure to obtain, is extracted 4 times with 8L water suspension medicinal extract, then with 8L ethyl acetate, ethyl acetate layer crosses silicon
Rubber column gel column, with methylene chloride-methanol system 50:1-1:2 gradient elution, collection 30:1-20:1 gradient elution object is fraction A, fraction A
Through ODS column chromatography, with ethanol-water system 30%, 60%, 90% elution collects 60% gradient elution object B, fraction B is through silica gel
Column obtains 4 fractions by methylene chloride-methanol system 50:1-1:2 gradient elution, washes what 20:1 and 10:1 gradient was collected
De- object is fraction C, and fraction C obtains 3 fractions through HP20 resin, with ethanol-water system gradient elution, is collected in 50% gradient
Eluate is fraction D, and fraction D collects fraction G, fraction H, fraction I, fraction by ODS column, through ethanol-water system elution respectively
J, fraction H are mainly collected in 40%, 45%, 50% gradient, and are prepared in 30%-45% acetonitrile-water by HPLC method
Compound 1, compound 2, compound 3, compound 4, compound 5.
The spectral information and nuclear magnetic signal ownership of five polyphenol compounds in 2 pithecellobium clypearia of embodiment
Compound 1:
Compound 1 is yellow oily compound (methanol), UV (MeOH) λ max (log ε): 200nm (2.42), 249nm
(3.27)；High-resolution electron spray ion massspectrum (HR-ESI-MS) provides quasi-molecular ion peak 349.0918 [M+H]+(calcd
for C17H17O8, 349.0918), in conjunction with1H-NMR,13C-NMR spectrum determines that molecular formula is C17H16O8, degree of unsaturation 10.In1H-NMR(400MHz,DMSO-d6, ppm, J in Hz) spectrum in, δH6.13 (1H, d, J=2.2, H-6), δH 6.20(1H,d,J
=2.2, H-8) be asymmetric four substituted benzene ring aromatic signal, δH7.05 (2H, s, H-3 ', H-7 ') are one
The aromatic signal of symmetrical four substituted benzene ring, δH3.38 (3H, s) are methoxy proton signal, δH5.15 (1H, t, J=
It 2.7, H-1) is time methylenedioxy group proton signal, δH 1.81(1H,m,H-2),δH1.97 (1H, m, H-2) are a mesomethylene carbon
On two together with even proton signal, δH 2.42(1H,m,H-3),δH2.58 (1H, m, H-3) are two on another group of mesomethylene carbon
It is a together with even proton signal, δH9.31 (4H, brs) are 4 phenolic hydroxyl group proton signals.In13C-NMR(100MHz,DMSO-d6,
Ppm 17 carbon signals, including 12 fragrant carbon signal δ) are provided in spectrumC 149.7(C-7),δC 152.7(C-5),δC 101.1
(C-8),δC 101.2(C-6),δC 107.3(C-4),δC 155.6(C-9),δC 118.5(C-2′),δC 109.0(C-3′,C-
7′),δC 145.7(C-4′,C-6′),δC139.2 (C-5 '), 2 sp3Hydridization carbon signal δC 25.2(C-2),δC 14.5(C-
3), 1 company oxygen carbon signal δC97.6 (C-1), 1 ester group carbon signal δC164.4 (C-1 ') and 1 methoxyl group carbon signal δC
55.3(-OCH3).In HMBC spectrum, H-1 is related to C-5, and H-2 is related to C-1, and H-3 is related to C-5, in conjunction with degree of unsaturation, mentions
Showing 4,5 of phenyl ring, there are benzene a pair of horses going side by side pyranoid ring segments.Methoxyl group hydrogen signal have to C-1 it is related, can determine methoxyl group connect
On 1.H-3 ', 7 ' have related, H-3 ' and C-5 ' to the ester carbonyl group of the position C-1 ', and 7 ' have correlation, H-7 ' and C-3 ', and 5 ' have phase
It closes, there are a galloyl segments in prompting structure.Compared with C-7 the even compound of hydroxyl, the compound C-7
Chemical displacement value is obviously mobile to High-Field, illustrates that galloyl is connected on the position C-7.H-6 and C-4,5,7,8 have a correlation, H-8 with
C-4,6,9 have correlation, it was demonstrated that another 1 hydroxyl is connected to 9, and the planar structure of the compound has been determined by analyzing above.Change
The absolute configuration for closing object 1 is obtained by comparing optical value and actual measurement optical value and calculating and actual measurement ECD is calculated, and works as C-1
When position absolute configuration is S, calculating optical value is(c 0.10,CH3OH), with actual measurement optical value (c
0.10,CH3OH) symbol is consistent, the Cotton effect peak in the experiment CD spectrum of compound 1 and the calculating ECD for being preset as 1S configuration
Cotton effect peak energy in spectrum is enough preferable identical, can determine that compound 1 is 1S configuration, and then the compound has been determined
Structure.Through scifinder database retrieval, compound 1 is the noval chemical compound that do not report.
Compound 2:
Compound 2 is buff oily compound (methanol), UV (MeOH) λ max (log ε): 210nm (2.78), 280nm
(2.97)；High-resolution electron spray ion massspectrum (HR-ESI-MS) provides quasi-molecular ion peak 383.0739 [M+Na]+(calcd
for C18H16NaO8, 383.0737), in conjunction with1H-NMR,13C-NMR spectrum determines that molecular formula is C18H16O8, degree of unsaturation 11.
In1H-NMR(400MHz,CD4O, ppm, J in Hz) spectrum in, δH6.22 (1H, d, J=2.3, H-8), δH 6.19(1H,d,J
=2.3, H-10) it is 2 aromatic signals on asymmetrical four substituted benzene ring, δH7.14 (2H, s, H-3 ', H-7 ') are pair
Claim 2 aromatic signals on four substituted benzene rings, δH4.52 (1H, dd, J=8.6,3.2, H-3) are the hydrogen of even oxygen methine
Signal, δH2.63 (2H, m, H-5) are a methene proton signal, δH 1.97(1H,m,H-4),δH 2.19(1H,m,H-4)
For the nonidentical methene proton signal of another magnetic, δH2.25 (3H, s, H-1) are 1 methyl proton signal.In13C-NMR
(100MHz,CD4O, ppm) 18 carbon signals are provided in spectrum, including 12 fragrant carbon signal δC108.3(C-6),δC 156.1(C-
7),δC 102.5(C-8),δC 102.1(C-10),δC 151.7(C-9),δC 157.4(C-11),δC 120.7(C-2′),δC
110.5(C-3′,C-7′),δC 146.7(C-4′,C-6′),δC140.5 (C-5 '), 1 ketone carbonyl carbon signal δC 210.0(C-
2), 1 company oxygen carbon signal δC81.5 (C-3), 3 sp3Hydridization carbon signal δC 19.0(C-5),δC 24.4(C-4),δC 26.2
(C-1), 1 ester group carbon signal δC167.0(C-1′).It is composed in conjunction with HMBC, H-1 and C-2, C-3 are related, prompt methyl and C-2
Carbonyl be connected, H-4 and C-2, C-5, C-6 are related, and H-5 and C-4, C-3, C-6, C-11 are related, H-3 and C-7, C-5, C-2, C-
4, it is related, it was demonstrated that 3 hydroxyls are connected with the position the C-7 of phenyl ring, form benzene a pair of horses going side by side pyranose ring structure, also prompt C-2 carbonyls and C-3
Position is connected.H-3 ' and C-7 ', C-5 ', C-2 ', C-1 ', C-4 ' are related, C-3 ' and C-7 ' can be speculated in phenyl ring in conjunction with carbon modal data
Upper in symmetric position, connect substituent, C-4 ', C-5 ', the position C-6 ' are not connected with hydroxyl group, and the position C-2 ' is connected with ester group,
Having prompted the compound, there are galloyl segments.H-10 is related to C-11, H-8 and C-6, and C-7, C-9 are related, composes in conjunction with carbon
C-11 are connected with hydroxyl group on Notes of Key Data phenyl ring, and the compound C-9 compared with C-9 the even compound of hydroxyl
Chemical shift is obviously mobile to High-Field, illustrates that galloyl is connected on the position C-9 of phenyl ring, so that it is determined that the putting down of the compound
Face structure.The absolute configuration of compound 2 is obtained by comparing calculating optical value and actual measurement optical value and calculating and actual measurement ECD
, when C-3 absolute configurations are S, calculating optical value is(c 0.10,CH3OH), with actual measurement optical value (c 0.10,CH3OH) symbol is consistent, Cotton effect peak in the experiment ECD of compound 2 spectrum and is preset as 3S configuration
The Cotton effect peak energy calculated in ECD spectrum is enough preferable identical, can determine that compound 2 is 3S configuration, and then this has been determined
The structure of compound.Through scifinder database retrieval, compound 2 is the noval chemical compound that do not report.
Compound 3:
Compound 3 is yellow oily compound (methanol), UV (MeOH) λ max (log ε): 207nm (2.36), 290nm
(3.48)；High-resolution electron spray ion massspectrum (HR-ESI-MS) provides quasi-molecular ion peak 481.0741 [M+Na]+(calcd
for C22H18NaO11, 481.0741), in conjunction with1H-NMR,13C-NMR spectrum determines that molecular formula is C22H18O11, degree of unsaturation 14.
In1H-NMR(400MHz,DMSO-d6, ppm, J in Hz) spectrum in, δH 6.98(2H,s,H-2″,H-6″),δH 7.05(2H,s,
H-3′,H-7′),δH6.12 (2H, s, H-6, H-8) are the 3 four aromatic signals replaced on symmetrical phenyl ring, δH 2.76
(2H, t, J=8.4, H-3), δH2.92 (2H, t, J=8.4, H-2) prompt for 2 methylene groups.In13C-NMR(100MHz,
DMSO-d6, ppm) and 22 carbon signals are provided in spectrum, including 18 fragrant carbon signal δC 107.6(C-2″,C-6″),δC 127.2
(C-1″),δC 138.8(C-4″),δC 145.6(C-3″,C-5″),δC 100.1(C-6,C-8),δC 111.4(C-4),δC149.4(C-7),δC 156.4(C-5,C-9),δC 109.0(C-3′,C-7′),δC 118.4(C-2′),δC 139.3(C-
5′),δC145.8 (C-4 ', C-6 '), 2 sp3Hydridization carbon signal δC 18.5(C-3),δC37.2 (C-2), 1 ester carbonyl group carbon
Signal δC164.5 (C-1 '), 1 ketone carbonyl carbon signal δC198.5(C-1).It is composed in conjunction with HMBC, H-3 ' and C-3 ', C-2 ', C-
4 ', C-1 ' are related, in combination with δC 109.0(C-3′,C-7′),δC 139.3(C-5′),δC145.8(C-4′,C-6′),δC
118.4 (C-2 ') prompt C-4 ' on phenyl ring, C-5 ' C-6 ' to be above connected with hydroxyl, unsubstituted on the position C-3 ', C-7 ', and C-2 ' is above connected with
Ester group can speculate that there are galloyl groups in structure.H-6 " with C-1 ", C-5 ", C-4 ", C-1, C-2 " are related,
In conjunction with carbon modal data δC 138.8(C-4″),δC 145.6(C-3″,C-5″),δC127.2 (C-1 ") are prompted, C-3 " on phenyl ring,
C-4 " C-5 " is connected with 3 phenolic hydroxyl groups, and C-1 carbonyls " are connected with C-1 on phenyl ring.H-2 and C-4, C-3, C-1 is related, H-3 and C-
4, C-1, C-9, C-2 are related, illustrate that C-4 are connected with 1 acyl ethyl segment.H-2 "/6 " and C-5 ", C-4 ", C-1 " are related, in conjunction with
δC 138.8(C-4″),δC 145.6(C-3″,C-5″),δC127.2 (C-1 ") illustrate that the position of substitution of three hydroxyls is C-3 ",
C-4 ", C-5 ", C-1 " are connected with substituent group, H-8 and C-4 on position, C-9, C-6, and C-7 is related, and C-5, C-9 chemical shifts to
Low field is mobile, can determine C-5 on phenyl ring, be connected with hydroxyl on C-9.The compound C-7 compared with C-7 the even compound of hydroxyl
Position chemical shift is obviously mobile to High-Field, illustrates to be connected with galloyl group on C-7, and then the knot of the compound has been determined
Structure.Through scifinder database retrieval, compound 3 is the noval chemical compound that do not report.
Compound 4:
Compound 4 is buff oily compound (methanol), UV (MeOH) λ max (log ε): 240nm (2.43), 280nm
(2.69)；High-resolution electron spray ion massspectrum (HR-ESI-MS) provides quasi-molecular ion peak 387.0674 [M+Na]+(calcd
for C17H16NaO9, 387.0687), in conjunction with1H-NMR,13C-NMR spectrum determines that molecular formula is C17H16O9, degree of unsaturation 10.
In1H-NMR(400MHz,DMSO-d6, ppm, J in Hz) spectrum in, δH 6.11(2H,s,H-6,H-8),δH 7.05(2H,s,H-
3 ', H-7 ') there are two symmetrical phenyl ring, δ in prompting structureH3.60 (3H, s) are methoxy proton signal, δH 2.41(2H,
T, J=8.0, H-2), δH2.76 (2H, t, J=8.0, H-3) are 2 methene proton signals, δH9.23 (5H, brs) are 5
Phenolic hydroxyl group proton signal.In13C-NMR(100MHz,DMSO-d6, ppm) spectrum in provide 17 carbon signals, including 12 aromatic carbons
Signal δC 110.6(C-4),δC 100.1(C-6,C-8),δC 156.5(C-5,C-9),δC 149.6(C-7),δC 118.5(C-
2′),δC 109.1(C-3′,C-7′),δC 145.9(C-4′,C-6′),δC139.3 (C-5 '), 2 sp3Hydridization carbon signal, δC
18.7(C-3),δC33.0 (C-2), 2 ester carbonyl group carbon signal δC 164.5(C-1′),δC173.3 (C-1), 1 methoxyl group carbon
Signal δC 51.3(-OCH3).It is composed in conjunction with HMBC, methoxyl group hydrogen signal is related to C-1, illustrates to be connected with 1 methoxyl group base on C-1
Group, H-2 and C-1, C-3, C-4 are related, and H-3 and C-1, C-2, C-4 are related, illustrate in compound there are an acyl propyl segment,
And it is connected with C-4 on phenyl ring.H-7 '/3 ' and C-1 ', C-2 ', C-3 ', C-5 ', C-6 ' are related, in combination with corresponding hydrogen
Spectrum, carbon modal data illustrate C-4 ', and the position C-5 ', C-6 ' is all connected with hydroxyl substituent, and the position C-2 ' is connected with ester group, thus suppositionization
There are a galloyl segments in conjunction object structure.H-6/8 and C-4, C-5, C-6, C-7, C-9 is related, composes in combination with carbon
On the position Notes of Key Data C-6, C-8 be not connected with substituent group, C-5, C-9 are all directly connected with hydroxyl group, and with C-7 companies
The compound of hydroxyl is obviously mobile to High-Field compared to C-7 chemical shifts of the compound, illustrates to be connected with galloyl on C-7
Group, and then the structure of the compound has been determined.Through scifinder database retrieval, compound 4 is the new chemical combination that do not report
Object.
Compound 5:
Compound 5 is yellow oily compound (methanol), UV (MeOH) λ max (log ε): 240nm (2,63), 281nm
(3.46)；High-resolution electron spray ion massspectrum (HR-ESI-MS) provides quasi-molecular ion peak 373.0532 [M+Na]+(calcd
for C16H14NaO9, 373.0530), in conjunction with1H-NMR,13C-NMR spectrum determines that molecular formula is C16H14O9, degree of unsaturation 10.
In1H-NMR(400MHz,DMSO-d6, ppm, J in Hz) spectrum in, δH7.05 (2H, s, H-3 ', H-7 ') prompt for symmetrical four and take
For 2 aromatic signals of phenyl ring, δH6.12 (2H, s, H-5, H-7) are 2 virtues on another symmetrical four substituted benzene ring
Fragrant proton signal.δH3.48 (2H, s, H-2) are methene proton signal, δH3.58 (3H, s) are methoxy proton signal.In13C-NMR(100MHz,DMSO-d6, ppm) and 16 carbon signals are provided in spectrum, including 12 fragrant carbon signal δC 105.8(C-3),
δC 156.7(C-4,C-8),δC 99.9(C-5,C-7),δC 150.2(C-6),δC 118.4(C-2′),δC 109.0(C-3′,
C-7′),δC 145.8(C-4′,C-6′),δC139.3 (C-5 '), 2 ester group signal δC164.4(C-1′),δC 171.8(C-
1), 1 sp3Hydridization carbon signal δC28.3 (C-2) and 1 methoxyl group carbon signal δC 51.4(-OCH3).It is composed in conjunction with HMBC, first
Oxygroup hydrogen signal is related to C-1, prompts to be connected with 1 methoxyl group on the position C-1.H-2 and C-8, C-3, C-4, C-1 are related, prompt
C-3 are connected with acyl methyl group on phenyl ring.H-7 ' and C-6 ', C-5 ', C-3 ', C-2 ', C-1 ' are related, in combination with δC 145.8
(C-4′,C-6′),δC139.3(C-5′),δC109.0 (C-3 ', C-7 ') prompt C-4 ' on phenyl ring, C-5 ' C-6 ' to be above connected with hydroxyl
Base, unsubstituted on the position C-3 ', C-7 ', C-2 ' are above connected with ester group, can speculate that there are galloyl groups.H-5 and
C-6, C-7, C-4, C-3 are related, in conjunction with δH6.12 (2H, s, H-5, H-7) and δC99.85 (C-5, C-7) speculate another benzene
Unsubstituted on the position C-5 on ring, C-7, C-4, C-8 chemical shifts it is mobile to low field, illustrate C-4 with C-8 on be connected with
Hydroxyl group, C-6 chemical shifts of the compound are obviously mobile to High-Field compared with C-6 the even compound of hydroxyl, illustrate C-6
It is connected with galloyl group on position, and then the structure of the compound has been determined.Through scifinder database retrieval, compound 5
For the noval chemical compound that do not report.
1 1~compound of compound 5 of table1H (100MHz) NMR data
2 1~compound of compound 5 of table13C (100MHz) NMR data
Five polyphenol compounds are in vitro to by H in 3 pithecellobium clypearia of embodiment2O2The neuroblastoma SH-SY5Y of induction
The neuroprotection of cellular damage measures
(1) H is established2O2The SH-SY5Y cellular damage model of induction
It chooses and is proliferated active SH-SY5Y cell in logarithmic growth phase, with 1.5 × 105The density of a/mL is seeded in 96
It is 37 DEG C in environmental condition in well culture plate, 5%CO2Incubator in after stationary culture 12h, observe the adherent situation of cell.It is empty
White control group adds not serum-containing medium, and H is not added in control group2O2, model group adds containing H2O2Culture solution without serum, is made into end
Concentration is 200 μm of ol/L, acts on 1,2,3,4,5,6h respectively, and above every group sets 3 multiple holes, surveys cell viability using MTT.
(2) compound is to H2O2Induce the protective effect of SH-SY5Y cellular damage
Model group, administration group, negative and 4 groups of blank control group, every group of 3 multiple holes are respectively set in experiment.
H2O2Model group: SH-SY5Y cell is successively through passing on, routine culture, after serum-free medium dilution, in 96 holes
It is inoculated in culture plate.Constant incubator (37 DEG C, 5%CO2) in stationary culture 12h to cell it is adherent after using contain serum free culture system
Liquid is then incubated for 2h, then adds H to every hole2O2Making final concentration of 200 μm of ol/L, MTT surveys cell viability after acting on 5h,
Administration group: cell culture is same as above, and is added contains untested compound (12.5,25,50 μm of ol/L) same item of culture solution later
Part is incubated for 2h, then adds H to each hole2O2(200 μm of ol/L) reacts 5h.
Negative control group: cell culture is same as above, and test-compound and H is not added2O2, other steps are same as above.
Blank control group: test-compound and H is not added2O2, inoculating cell, other steps are not same as above.
Every hole absorbance value (A) finally is surveyed with microplate reader (λ=490nm), calculates survival rate, formula is as follows:
(3) experimental result:
To the anti-H of 5 compounds2O2The protective effect of the SH-SY5Y cellular damage of induction is screened, as a result as shown in the table:
Cell survival rate after table 3 is administered
The above results show that compound 1 has preferable anti-H at 12.5 μM, 25 μM, 50 μM2O2The neuroprotection of induction
Activity, cell survival rate are higher than 30% or more model group at 12.5 μM, are higher than 40% or more model group at 25 μM, in 50 μM of height
In 50% or more model group.Compound 2 has preferable anti-H at 50 μM2O2The neuroprotective activity of induction, cell survival
Rate is above 40% or more model group.Compound 3 has preferable anti-H at 25 μM, 50 μM2O2The neuroprotective activity of induction,
Its cell survival rate is being higher than 10% or more model group at 25 μM, is higher than 30% or more model group at 50 μM.Compound 4 is at 50 μM
When there is preferable anti-H2O2The neuroprotective activity of induction, cell survival rate are above 10% or more model group.Compound 1,
Compound 2, compound 3, compound 4 have the anti-H for being better than positive drug in various concentration2O2The neuroprotective activity of induction, chemical combination
Object 5 has and the comparable anti-H of positive drug2O2The neuroprotective activity of induction.
Claims (9)
1. such as flowering structure compound represented:
2. the preparation method of compound as described in claim 1, which comprises the following steps:
(1) dry pithecellobium clypearia branches and leaves are taken, are extracted with 70-80% ethyl alcohol, extracting solution is obtained and is condensed into medicinal extract；Total medicinal extract water
It after suspension, is extracted with ethyl acetate, ethyl acetate layer uses silicagel column, carries out gradient with methylene chloride-methanol system and washes
It is de-, collect fraction A；
(2) fraction A is eluted through ODS column chromatography through ethanol-water system, collects fraction B；
(3) fraction B collects fraction C by methylene chloride-methanol system elutions through silicagel column；
(4) fraction C collects fraction D through HP20 resin with ethanol-water system gradient elution；
(5) fraction D collects fraction G, fraction H, fraction I, fraction J, fraction H through ethanol-water system elution by ODS column respectively
It is mainly collected in 40%-50% gradient, and compound 1, compound 2, compound is prepared in acetonitrile-water by HPLC method
3, compound 4, compound 5.
3. the preparation method of compound as claimed in claim 2, which is characterized in that dichloromethane in step (1) or step (3)
Alkane-methanol gradient are as follows: 50:1-1:2.
4. the preparation method of compound as claimed in claim 2, which is characterized in that alcohol-water in step (2) or step (4)
Gradient be 30%-90%.
5. the preparation method of compound as claimed in claim 2, which is characterized in that the volume ratio of acetonitrile-water in step (5)
Are as follows: 30%-45%.
6. pharmaceutical composition includes one or more of compound described in claim 1 and pharmaceutically acceptable carrier
Or excipient.
7. application of the compound described in claim 1 in preparation prevention or treatment neurodegenerative disease drug.
8. application of the pharmaceutical composition as claimed in claim 6 in preparation prevention or treatment neurodegenerative disease drug.
9. application as claimed in claim 7 or 8, which is characterized in that the neurodegenerative disease is H2O2Induce SH-
Neurodegenerative disease caused by SYSY cellular damage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910778418.8A CN110483462B (en) | 2019-08-22 | 2019-08-22 | Polyphenol compound with neuroprotective activity and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910778418.8A CN110483462B (en) | 2019-08-22 | 2019-08-22 | Polyphenol compound with neuroprotective activity and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110483462A true CN110483462A (en) | 2019-11-22 |
| CN110483462B CN110483462B (en) | 2021-06-18 |
Family
ID=68552876
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910778418.8A Active CN110483462B (en) | 2019-08-22 | 2019-08-22 | Polyphenol compound with neuroprotective activity and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110483462B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104974018A (en) * | 2015-06-15 | 2015-10-14 | 沈阳药科大学 | Compounds extracted from traditional Chinese medicine Pithecellobium clypearia Benth. and use thereof |
| CN108610387A (en) * | 2018-03-19 | 2018-10-02 | 沈阳化工大学 | One kind having active four isoflavan glycosides compounds of neurocyte protection and preparation method thereof |
-
2019
- 2019-08-22 CN CN201910778418.8A patent/CN110483462B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104974018A (en) * | 2015-06-15 | 2015-10-14 | 沈阳药科大学 | Compounds extracted from traditional Chinese medicine Pithecellobium clypearia Benth. and use thereof |
| CN108610387A (en) * | 2018-03-19 | 2018-10-02 | 沈阳化工大学 | One kind having active four isoflavan glycosides compounds of neurocyte protection and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| WANG YU-XI等: "Enantiomeric lignans with anti-β-amyloid aggregation activity from the twigs and leaves of Pithecellobium clypearia Benth", 《BIOORGANIC CHEMISTRY》 * |
| WANG YU-XI等: "Flavonoids and their derivatives with b-amyloid aggregation inhibitory activity from the leaves and twigs of Pithecellobium clypearia Benth", 《BIOORGANIC&MEDICINAL CHEMISTRY LETTERS》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110483462B (en) | 2021-06-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104151373B (en) | Lignan glycoside compounds and preparation method thereof | |
| Jin et al. | Four new compounds from the bulbs of Lycoris aurea with neuroprotective effects against CoCl 2 and H 2 O 2-induced SH-SY5Y cell injuries | |
| CN112159378B (en) | Gemmaxane type sesquiterpene lactone compound and preparation method and application thereof | |
| CN103819445A (en) | Method for preparing two neo-pentenyl flavonoid compounds with hypolipidemic activity in fructus podophylli | |
| CN110452113A (en) | A kind of (4 → 2) reset Crow alkane type diterpene-kind compound and its preparation method and application | |
| CN108191931B (en) | Three new compounds in Xinjiang peony and health care and medical application thereof | |
| CN110305014B (en) | Cycloneolignanoid enantiomers, preparation and application thereof | |
| CN111253247B (en) | Preparation method and application of novel phenolic acid compound with anti-inflammatory activity | |
| CN115108935B (en) | Alkaloid compound in white English and preparation method and application thereof | |
| CN104292203A (en) | Isocoumarin compound and preparation method and application thereof | |
| CN110511194A (en) | Tetrahydrofuran type lignan compound and its preparation method and application | |
| He et al. | Antitrichomonas in vitro activity of Cussonia holstii Engl | |
| CN115991692A (en) | Preparation method and application of spirodienone lignan compound in Isatis tinctoria | |
| CN110357894A (en) | A kind of tricyclic alkaloid compound and the preparation method and application thereof | |
| CN110483462A (en) | Polyphenol compound and preparation method thereof with neuroprotective activity | |
| CN110551163A (en) | Method for extracting flavonoid glycoside compounds from fructus Podophylli and application thereof | |
| CN106674086B (en) | A kind of piperidones Alkaloid compound and its preparation method and application | |
| CN113105471B (en) | Phenolic acid compound with antioxidant activity and application thereof | |
| CN108948040B (en) | Gilmaxane type sesquiterpene compound extracted from herba Centellae and application thereof | |
| CN109734696A (en) | A kind of new diepoxy lignan compound and preparation method thereof | |
| CN106565444A (en) | Extraction method and application of phenanthrene compounds from overground part of Chinese yam | |
| CN116768912B (en) | Spiroindolone alkaloid compound in Isatis tinctoria as well as preparation method and application thereof | |
| CN105669692B (en) | A kind of extracting method and purposes of phthalide-type dimer compound | |
| CN111484485A (en) | Anti-inflammatory macrocyclic polyamine alkaloid celacarfurine | |
| CN116903578B (en) | Phenolic acid compound in Glechoma hederacea as well as extraction and separation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |